Essential role of tubulin-folding cofactor D in microtubule assembly and its association with microtubules in fission yeast.
The main structural components of microtubules are alpha- and beta-tubulins. A group of proteins called cofactors are crucial in the formation of assembly-competent tubulin molecules in vitro. Whilst an in vitro role is emerging for these cofactors, their biological functions in vivo remain to be established. In order to understand the fundamental mechanisms that determine cell polarity, we have screened for fission yeast mutants with altered polarity. Here we show that alp1+ encodes a homologue of cofactor D and executes a function essential for cell viability. A temperature-sensitive alp1 mutant shows a variety of defects including abnormal mitoses, loss of microtubule structures, displacement of the nucleus, altered growth polarity and asymmetrical cell division. Overexpression of Alp1 is lethal in wild-type cells, resulting in altered cell shape, but is rescued by co-overexpression of beta-tubulin. Alp1 co-localizes with microtubules, both interphase arrays and mitotic spindles. Furthermore, Alp1 binds to and co-sediments with taxol (paclitaxel)-stabilized porcine microtubules. Our results suggest that, in addition to a function in the folding of beta-tubulin, cofactor D may play a vital role in microtubule-dependent processes as a microtubule-associated protein.
Hirata
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D
Laboratory of Cell Regulation, Imperial Cancer Research Fund, PO Box 123, 44 Lincoln's Inn Fields, London WC2A 3PX, UK.
Masuda
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Eddison
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M
Toda
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T
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000378:metabolism / Q000648:ultrastructure; D016764:Cell Polarity / Q000235:genetics / Q000502:physiology; D005656:Fungal Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D015870:Gene Expression / Q000235:genetics / Q000502:physiology; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000502:physiology*; D008870:Microtubules / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics / Q000502:physiology; D011487:Protein Conformation; D017510:Protein Folding; D012568:Schizosaccharomyces / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D029702:Schizosaccharomyces pombe Proteins; D017386:Sequence Homology, Amino Acid; D013552:Swine; D013696:Temperature; D014404:Tubulin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014461:Ultracentrifugation
C110653:Alp1 protein, S pombe; D005656:Fungal Proteins; D008869:Microtubule-Associated Proteins; D029702:Schizosaccharomyces pombe Proteins; D014404:Tubulin